ChIP-Atlas: SRX5352439

Antigen

Cell type Class: Gonad
Cell type: Spermatogonia
MeSH Description: Euploid male germ cells of an early stage of SPERMATOGENESIS, derived from prespermatogonia. With the onset of puberty, spermatogonia at the basement membrane of the seminiferous tubule proliferate by mitotic then meiotic divisions and give rise to the haploid SPERMATOCYTES.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Testes were removed and decapsulated. They were then digested with collagenase B (2mg/mL) for 10 minutes at 37°C. After rinsing in PBS, another digestion with 0.25% trypsin-EDTA for 5 minutes at 37°C was performed. The resulting cell suspension was washed twice, resuspended in sterile phosphate buffer solution (PBS) with DNase (50μg/mL) and filtered (40μm). Spermatogonia were isolated by fluorescence-activated cell sorting of Oct4-GFP positive germ cells with a FACSAria III laser sorter. ChIP-seq libraries were prepared using the Kapa HTP Library Preparation Kit (Roche) as per the manufacturer's recommendations except that adapters and PCR primers were diluted 100-fold and the size selection step was done after the PCR step.